High-resolution deconvolution of selection effects on the composition of the B cell repertoire

项目摘要 V（D）J重组的随机性质，其主要目的是产生结合广泛的抗体。 一系列潜在的目标，往往无意中产生抗体，识别自身抗原。等 自身反应性抗体通常与自身免疫性疾病相关， 数百万人，对社会构成重大财政负担。在早期发育过程中， 用于管理自身反应性B细胞，包括缺失、修饰或沉默。在外周，B细胞 正在经历亲和力成熟的人受到外周耐受机制的影响，以防止获得高亲和力。 亲和自身反应性。事实上，无数的选择和容忍机制，跨越整个B 从早期的前B细胞到长寿的浆细胞的细胞谱系，对细胞的组成产生巨大的影响。 人抗体库。尽管人们认为选择和宽容的剧目塑造效应 尽管如此，我们对B细胞库组成的方式仍然只有有限的了解， 通过选择来调节。 我实验室的长期研究重点是利用高通量测序技术， 了解人类B细胞库的发育、成熟和功能。继续 技术进步，包括新兴的单细胞分析技术，允许构建 越来越详细的分子概况为大量的个别细胞。超高分辨率的 这样的数据集将允许在前所未有的深度和细节水平上研究体液免疫系统。 在接下来的五年里，我们将利用这些进步来解决 以下地区。(1)发现由B细胞受体编码的特定特征或特征模式， 通过中枢耐受性选择性地耗尽或修改。(2)早期B细胞发育检查点的鉴定 选择和宽容使不同个体的所有技能都同质化。(3)表征 亲和力成熟的全局模式，其独立于任何特定的抗原，全局地塑造 记忆库的组成。更全面地了解过程和机制， 塑造B细胞库是至关重要的，并与正在进行的感染性疾病研究广泛相关， 自身免疫和合理的疫苗开发。

Project Summary The random nature of V(D)J recombination, whose primary purpose is to produce antibodies that bind a broad range of potential targets, often inadvertently creates antibodies that recognize self antigens. Such autoreactive antibodies are often associated with autoimmune diseases that adversely affect the wellbeing of millions and represent a significant financial burden on society. During early development, mechanisms exist for managing autoreactive B cells, including deletion, modification, or silencing. In the periphery, B cells undergoing affinity maturation are subject to peripheral tolerance mechanisms to prevent acquisition of high affinity autoreactivity. Indeed, a myriad of selection and tolerance mechanisms, spanning the entirety of the B cell lineage from early pro-B cells to long-lived plasma cells, exert massive influence on the composition of the human antibody repertoire. Although the repertoire-shaping effects of selection and tolerance are thought to be quite large, we still have only a limited understanding of the ways in which B cell repertoire composition is regulated by selection. The long-term research focus of my laboratory is to use high-throughput sequencing to gain a more complete understanding of the development, maturation and function of the human B cell repertoire. Continuing technological advances, including emerging single-cell analysis techniques, allow the construction of increasingly detailed molecular profiles for large numbers of individual cells. The extremely high resolution of such datasets will allow study of the humoral immune system at an unprecedented level of depth and detail. Over the next five years, we will leverage these advances to address significant knowledge gaps in the following areas. (1) Discovery of specific features or feature patterns encoded by B cell receptors that are selectively depleted or modified by central tolerance. (2) Identification of early B cell development checkpoints at which selection and tolerance homogenize the repertoires of different individuals. (3) Characterization of global patterns of affinity maturation which, independent of any particular antigen, globally shape the composition of the memory repertoire. A more complete understanding of the processes and mechanisms that shape the B cell repertoire is vitally important and broadly relevant to ongoing research in infectious disease, autoimmunity, and rational vaccine development.